山东大学耳鼻喉眼学报 ›› 2018, Vol. 32 ›› Issue (2): 84-87.doi: 10.6040/j.issn.1673-3770.0.2017.323

• ·论著· • 上一篇    下一篇

白花蛇舌草多糖提取物诱导Hep-2细胞凋亡与抑制侵袭机制

赵小燕,吴彩琴,任晓勇,王正辉   

  1. 2015SF044)第一作者:赵小燕。 E-mail:632453562@qq.com通讯作者:王正辉。 E-mail:ehui4298@163.com DOI:10.6040/j.issn.1673-3770.0.2017.323白花蛇舌草多糖提取物诱导Hep-2细胞凋亡与抑制侵袭机制赵小燕, 吴彩琴, 任晓勇, 王正辉(西安交通大学第二附属医院耳鼻咽喉-头颈外科, 陕西 西安 710004
  • 收稿日期:2017-07-28 出版日期:2018-03-20 发布日期:2018-03-20
  • 通讯作者: 王正辉. E-mail:ehui4298@163.com
  • 基金资助:
    陕西省科技攻关项目(2015SF044)

A polysaccharide from hedyotis diffusa induces apoptosis and inhibits migration in Hep-2 cells

ZHAO Xiaoyan, WU Caiqin, REN Xiaoyong, WANG Zhenghui   

  1. Department of Otolaryngology-Head and Neck Surgery, The Second Affiliated Hospital, Xian Jiaotong University, Xian 710004, Shaanxi, China
  • Received:2017-07-28 Online:2018-03-20 Published:2018-03-20

摘要: 目的 研究白花蛇舌草多糖提取物(HDP)对人喉癌Hep-2细胞的抗瘤作用。 方法 采用MTT测量细胞活性,流式细胞仪进行凋亡分析。Transwell培养小室进行细胞侵袭实验,Western blotting测量多糖提取物诱导凋亡和抗侵袭活性的机制。 结果 HDP对喉癌细胞的抑制呈时间和浓度依赖性,浓度越高,时间越长抑制率越高。细胞周期分析发现HDP(400 μg/mL)使细胞滞留于G0/G1期,同时发现药物作用24 h后细胞凋亡率明显升高,Western blotting结果分析发现相关凋亡蛋白caspase-3, caspase-8和caspase-9表达升高,而Bcl-2表达下降。同时细胞侵袭实验还发现HDP抑制喉癌细胞的侵袭,抑制MMP-2和uPA相关蛋白的表达。 结论 研究发现 HDP 抑制喉癌细胞的凋亡和侵袭,对恶性肿瘤的治疗有潜在的应用价值。

关键词: 白花蛇舌草, Hep-2细胞, 侵袭, 凋亡, 多糖

Abstract: Objective The aim of this study was to explore the anti-tumor potential of a polysaccharide isolated from Hedyotis diffusa(HDP)in Hep-2 human larynx squamous carcinoma cells. Methods Cell cycle and apoptosis were analyzed by detecting changes in protein expression by flow cytometry. Apoptosis was further characterized by detecting relevant proteins by western blotting. Results HDP suppressed the proliferation of Hep-2 cells in a time- and dose-dependent manner. Cell cycle analysis revealed that exposure to HDP(400 μg/mL)caused G0/G1 cell cycle arrest in Hep-2 cells. Moreover, flow cytometry revealed that Hep-2 cells treated with HDP for 24 h underwent apoptosis to a significantly greater extent than untreated control cells. HDP treatment promoted the expression of caspase-3, caspase-8, and caspase-9, and reduced the expression of Bcl-2. HDP also inhibited cell migration and suppressed the expression of MMP-2 and uPA. Conclusion HDP possesses suppressive effects against aggressive phenotypes of human larynx squamous carcinoma cells and may have therapeutic potential for this particular malignancy.

Key words: Hedyotis diffusa, Polysaccharide, Hep-2 Cells, Apoptosis, Migration

中图分类号: 

  • R739.6
[1] Hu E, Wang D, Chen J, et al. Novel cyclotides from hedyotis diffusa induce apoptosis and inhibit proliferation and migration of prostate cancer cells[J]. Int J Clin Exp Med, 2015, 8(3):4059-4065.
[2] Lin JM, Wei LH, Xu W, et al. Effect of hedyotis diffusa willd extract on tumor angiogenesis[J]. Mol Med Rep, 2011, 4(6):1283-1288.
[3] 苏芳, 魏菁, 何龙. 白花蛇舌草对人喉癌细胞凋亡及增殖的影响[J]. 实用中医内科杂志, 2012, 26(7):1-2. SU Fang, WEI Jing, HE Long. The effect of herba hedyotis diffusae on apoptosis and proliferation of human laryngeal carcinoma cells[J]. J Prac Trad Chin Int Med, 2012, 26(7):1-2.
[4] 张军, 张耀明, 王正辉, 等. 草苁蓉多糖提取物诱导人喉癌Hep2细胞凋亡的实验研究[J]. 陕西医学杂志, 2014, 43(8):947-949. ZHANG Jun, ZHANG Yaoming, WANG Zhenghui, et al. Polysaccharide of boschniakia rossica induces apoptosis on Hep2 cells[J]. Shaanxi Med J, 2014, 43(8):947-949.
[5] 张健梅, 文姝, 李洁, 等. 三氧化二砷与紫杉醇联合诱导Hep-2细胞凋亡的体外研究[J]. 中国耳鼻咽喉头颈外科, 2011, 18(4):196-199. ZHANG Jianmei, WEN Shu, LI Jie, et al. Study in the effect of arsenic trioxide and taxol on Hep-2 in vitro[J]. Chin Arch Otolaryngol Head Neck Surg, 2011, 18(4):196-199.
[6] Wang SY, Hsu ML, Hsu HC, et al. The anti-tumor effect of ganoderma lucidum is mediated by cytokines released from activated macrophages and T lymphocytes[J]. Int J Cancer, 1997, 70(6):699-705.
[7] Wasser SP. Medicinal mushrooms as a source of antitumor and immunomodulating polysaccharides[J]. Appl Microbiol Biotechnol, 2002, 60(3):258-274.
[8] Zhang P, Zhang B, Gu J, et al. The study of the effect of hedyotis diffusa on the proliferation and the apoptosis of the cervical tumor in nude mouse model[J]. Cell Biochem Biophys, 2015, 72(3):783-789.
[9] Zhang Y, Xie RF, Xiao QG, et al. Hedyotis diffusa willd extract inhibits the growth of human glioblastoma cells by inducing mitochondrial apoptosis via AKT/ERK pathways[J]. J Ethnopharmacol, 2014, 158:404-411.
[10] Lu QL, Poulsom R, Wong L, et al. Bcl-2 expression in adult and embryonic non-haematopoietic tissues[J]. J Pathol, 1993, 169(4):431-437.
[11] Cui M, Zhang M, Liu HF, et al. Effects of microRNA-21 targeting PITX2 on proliferation and apoptosis of pituitary tumor cells[J]. Eur Rev Med Pharmacol Sci, 2017, 21(13):2995-3004.
[12] Garner TP, Lopez A, Reyna DE, et al. Progress in targeting the BCL-2 family of proteins[J]. Curr Opin Chem Biol, 2017, 39:133-142.
[13] 孙翔宇, 何丽娜, 吕克文, 等. 三氧化二砷对人舌鳞癌细胞CAL-27侵袭能力的影响[J]. 国际口腔医学杂志, 2014, 41(5):541-545. SUN Xiangyu, HE Lina, LÜ Kewen, et al. The effect of arsenic trioxide on invasion of human tongue squamous carcinoma cell line CAL-27[J]. Inter J Stom, 2014, 41(5):541-545.
[14] Kawata R, Shimada T, Maruyama S, et al. Enhanced production of matrix metalloproteinase-2 in human head and neck carcinomas is correlated with lymph node metastasis[J]. Acta Otolaryngol, 2002, 122(1):101-106.
[15] 刘斐斐, 李薇, 尚伟, 等. 喉鳞癌组织MMP-2和uPA表达与肿瘤侵袭转移关系[J]. 青岛大学医学院学报, 2008, 44(3):238-240. LIU Feifei, LI Wei, SHANG Wei, et al. Expressions of MMP-2 and uPA in laryngeal squamous cell carcinoma and their relationship with lymph node metastasis[J]. Act Acad Med Qingdao Univ, 2008, 44(3):238-240.
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